骨髓间充质干细胞移植下调MODS兔心、脾、肾组织中细胞凋亡因子Apaf-1 mRNA表达  

Bone Marrow Mesenchymal Stem Cells Transplantation Down- regulates the mRNA Expression of the Apoptotic Factor Apaf-1 in Hearts,Spleens,and Kidneys of Rabbits with MODS

作　　者：凌斌[1] 陈静[1] 王廷华[2] 潘兴华[3] 孙洁[1] 

机构地区：[1]云南省第二人民医院ICU科,云南昆明650031 [2]昆明医学院神经科学研究所,云南昆明650031 [3]中国人民解放军成都军区昆明总医院临床实验科,云南昆明650031

出　　处：《昆明医学院学报》2010年第3期14-20,共7页Journal of Kunming Medical College

基　　金：云南省自然科学基金资助项目(2007C249M)

摘　　要：目的观察骨髓间充质干细胞(MSC)移植对多器官功能不全综合症(MODS)兔受累器官组织细胞凋亡蛋白酶活化因子-1 mRNA(Apaf-1 mRNA)表达及影响,探讨应用于MODS治疗的可能性和应用前景.方法体外培养、扩增、鉴定兔MSC,慢病毒转基因标记GFP,经股动脉移植,观察MSC移植后,在MODS动物模型宿主内的存活情况.RT-PCR测定MSC移植前后Apaf-1 mRNA在心、脾、肾脏器中的表达变化.结果光镜观察MSC符合MSC细胞形态学的特征;激光共聚焦显微镜观察,移植MSC兔的心、脾、肾脏器组织内有慢病毒转基因标记GFP的MSC存在;流式细胞检测分析MSC不表达血细胞表面标记如CD34、CD45,但表达CD73、CD105;RT-PCR检测结果显示正常心、脾、肾组织中均有Apaf-1 mRNA的表达,模型组心、脾、肾Apaf-1 mRNA的表达较正常组明显增加(P<0.01),而移植组Apaf-1 mRNA的表达低于模型组(P<0.01).结论体外培养获得的兔MSC表现MSC形态学特征;移植后的MSC能够定植在MODS兔的受损脏器组织内;MSC能够下调Apaf-1 mRNA在MODS的心、脾、肾组织中的表达,提示MSC移植有望为MODS提供新的治疗方法.Objective To observe the effect of bone marrowmesenchymal stem cells transplantation on the mRNA expression of the apoptotic factor Apaf-1 in involved organs of rabbits with MODS,and explore its possibility and potential in the treatment of MODS.Methods Bone marrow mesenchymal stem cells（MSCs） of rabbits were cultured,expanded,and indentified in vitro,and were marked by lentivirus transgenic marker GFP.MSCs were transplanted through the femoral artery to rabbits with MODS,and then the survival of MSCs in rabbits was observed.RT-PCR was used to detected the mRNA expression of Apaf-1 in hearts,spleens,and kidneys of rabbits before and after MSCs transplantation.Results The morphology of MSCs under light microscopy was in line with the morphological characteristics of MSCs.MSCs marked by lentivirus transgenic marker GFP were found in hearts,spleens,and kidneys of rabbits under laser confocal microscope.The flow cytometry analysis results showed that CD34 and CD45 were not expressed while CD73 and CD105 were expressed in MSCs.RT-PCR re-sults showed Apaf-1 mRNA was expressed in hearts,spleens,and kidneys of healthy rabbits in control group,the Apaf-1 mRNA expression significantly increased in these oragans of rabbits with MODS in model group（P 0.01）,and the Apaf-1 mRNA expression was lower in transplantation group than in model group（P0.01）.Conclusions The rabbit MSCs cultured in vitro showthe morphological characteristics of MSCs.MSCs can col-onize and survive in the damaged organs of rabbits with MODS after transplantation.MSCs can down-regulate the mRNA expression of the apoptotic factor Apaf-1 in involved organs of rabbits with MODS,suggest that MSCs transplantation may be a potential treatment method for MODS.

关 键 词：骨髓间充质干细胞 多器官功能不全综合征 GFP标记 兔 移植 细胞凋亡 APAF-1 

分 类 号：R614[医药卫生—麻醉学]